KR20070109089A - Anticancer agent with momilacton b and rice hull as active ingredient from rice hulls - Google Patents

Abstract

A composition comprising an anti-cancer active material of momilactone B isolated from rice hulls is provided to inhibit the growth of colon cancer cell line without influencing on normal cells. An anticancer composition comprises momilactone B as an effective ingredient, wherein the momilactone B is extract from rice hull using an organic solvent such as methanol and ethanol. The composition further comprises a pharmaceutically acceptable carrier such as saline solution, a buffering agent, dextrose, water, glycerol, linger solution, lactose, sucrose, calcium silicate, methyl cellulose and ethanol.

Description

Anticancer composition containing chaff and momilactone {as an active ingredient {Anticancer agent with momilacton B and rice hull as active ingredient from rice hulls}

1 is a graph showing the cell growth inhibition of colon cancer cell line HT-29 of chaff extract according to the present invention.

Figure 2 is a graph showing the ¹ H NMR spectrum (A) and ¹³ C NMR spectrum (B) by the ^first NMR measurement of anti-cancer substances in chaff extract of the present invention.

Figure 3 is a graph showing the spectrum of HMBC (A), HMQC (B) and COSY (C) by the first NMR measurement of anti-cancer substances in chaff extract of the present invention.

Figure 4 is a graph showing the cell growth inhibition of 24 hours after (A), 48 hours (B) and 72 hours (C) after treatment of the mother cancer lactone B HT-29 and SW620 of the present invention.

FIG. 5 is a graph showing the inhibition of colony formation after treatment of Momiractone B of the present invention to colon cancer cell lines HT-29 and SW620. FIG.

Figure 6 is a graph showing the cell growth inhibition ability of the mother cell lactone B of the present invention to the normal cell line CCD986sk.

The present invention relates to a composition for anticancer containing chaff and momi lactone B as an active ingredient, more specifically, chaff which is an agricultural processing by-product extracted with alcohol, and after separating the momi lactone B from it, the added value is It is about developing into a high anticancer composition.

Recently, as cancer chemotherapy is advanced, new anticancer drugs or treatments have been developed, and treatment or extension of life expectancy is expected, and in particular, some cancers such as leukemia, malignant lymphoma, and adrenal gland cancer are getting high therapeutic effects. On the other hand, resistant cancers resistant to anticancer drugs appear during treatment, and solid cancers representing gastrointestinal cancers that hardly respond to anticancer drugs are present, and cancer is still the leading cause of death in developed countries. Among them, new anticancer drugs such as irinodecane hydrochloride (CPT-II) and paclitaxel (taxol) can be expected to some extent not only for blood cancer but also for some solid cancers. The problem of selectivity still remains a problem. Among them, the biggest problem of the anticancer drugs used in the clinic can be said that the biggest problem is the inability to selectively distinguish between cancer cells and normal cells. In order to solve this problem, studies on cancer biology from various angles have been conducted to study the characteristics of cancer at the molecular level such as cell cancer, drug resistance or metastasis. Successful examples of cancer molecule targeting agents include Gleevec, a Bcr-Abl inhibitor, and Iressa, an inhibitor of EGF receptor tyrosine kinase.

Currently, the development of anticancer drugs centered on natural products has been applied to extracts of plants, which have traditionally been used as folk remedies for a long time, for cancer genes, cancer suppressor genes, signal transduction factors, and drug resistance susceptibility factors established by the development of modern cell biological research. Screening around cancer molecular targets, such as apoptosis-related factors, can be said to develop an anticancer agent that separates and purifies active substances and determines the structure of the active substances. In particular, rice is an important grain that is used as a staple food in Asia, Africa and Latin America, and is produced in Korea by more than 5.3 million tons per year. The rice husk generated during rice milling accounts for 18-20% of rice production. It is estimated to generate more than 1 million tons per year.

Recent research has shown that rice husks generated during the milling of rice have many antioxidants and biologically active substances naturally to combat the oxidative effects of the external environment. It is being done.

Therefore, the present inventors have separated the new high value-added anticancer active substance from methanol extract of rice husk, an unused agricultural processing product that is discarded, and examined the chemical structure and physicochemical characteristics, and thus the compound is called momilactone B and biological activity. As a result, the present invention was found to exhibit activity inhibiting the growth of colorectal cancer cell lines, thereby completing the present invention.

Accordingly, an object of the present invention is to provide a novel use compound having excellent activity of inhibiting the growth of colorectal cancer cell lines from extracts based on chaff which is an unused resource.

It is another object of the present invention to provide an extract that contains an active ingredient that inhibits the growth of the powerful colorectal cancer cell line and an effective ingredient that does not affect normal cells.

In order to achieve the above object, the present invention provides an anticancer composition containing Momilatone B as an active ingredient.

Hereinafter, the present invention will be described in detail.

At this time, if there is no other definitions in the technical terms and scientific terms used herein, those having ordinary skill in the art to which this invention belongs have the meaning that is commonly understood.

In addition, repeated description of the same technical configuration and operation as in the prior art will be omitted.

Rice hulls in the present invention refers to the outer shell of rice.

The present invention is to enable the development of an anticancer composition for treating cancer diseases using Momiractone B.

In addition, the present invention enables the extraction of Momilactone B from the almost discarded rice husk.

In this invention, it is preferable to extract Momilactone B from rice hulls from an organic solvent. Specifically, alcohols such as methanol or ethanol can be used.

Therefore, the present invention more specifically provides an anticancer composition containing Momilatone B as an active ingredient.

Specifically, the present invention, in order to extract momilactone B, which is one of the anticancer components, from chaff, first extracting chaff into alcohol; The methanol extract was separated and extracted with ethyl acetate, separated by silica gel column chromatography, Sephadex LH-20 column chromatography, reverse phase TLC, and then HPLC was performed under an acetonitrile solvent to give a compound of the present invention momilactone B (momilactone B). Purification); It includes.

However, chaff extract and momi lactone B compound of the present invention is not limited by the above-described method of separation and extraction from chaff, it can be obtained by combining other known methods alone or suitably.

In addition, as disclosed in the Remington's Pharmaceutical Sciences Handbook, Mack Pub, Co., NY, USA, by using Momilactone B of the present invention as an active ingredient, depending on the administration method, dosage form and therapeutic purpose The mixture is diluted with a pharmaceutically acceptable carrier in the form of a suitable pharmaceutical composition or enclosed in a carrier in a container form for use.

When the carrier is used as a diluent, saline, buffer, dextrose, water, glycerol, Ringer's solution, lactose, sucrose, calcium silicate, methyl cellulose or ethanol are used for oral administration and parenteral administration using the carrier. It is prepared in the form of powders, granules, injection solutions, syrups, solutions, tablets, suppositories, and the like. However, the carrier of the present invention is not limited to the above carrier.

In this case, parenteral administration refers to the administration of the drug through rectal, intravenous, peritoneal, muscle, arterial, transdermal, nasal, inhalation, in addition to oral.

In addition, the formulation may further include fillers, anti-coagulants, lubricants, wetting agents, flavors, emulsifiers, preservatives, and the like, and may be formulated to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal.

On the other hand, the effective amount of momilactone B in the anticancer composition of the present invention will be determined by those skilled in the art depending on the type of cancer, the route of administration, the dosage form, the purpose of use, and the severity of the disease of the patient.

Hereinafter, the present invention will be described in more detail with reference to specific examples. However, the present invention is not limited to the following examples, and it will be apparent to those skilled in the art that various changes or modifications can be made within the spirit and scope of the present invention.

Example 1 Growth Inhibition Activity of Colorectal Cancer Cell Line from Chaff Methanol Extract of the Present Invention

In this example, to reduce the growth inhibition of the colon cancer cell line of the chaff methanol extract of the present invention was subjected to an MTT reduction assay (MTT reduction assay) using a human-derived colorectal cancer cell line HT-29. The cells were cultured in an incubator at 37 ° C. and 5% CO ₂ using RPMI1640 medium, and then incubated for 24 hours in a concentration of 1.5 × 10 ⁴ cells / well in a 96-well plate, followed by treatment with the rice husk methanol extract of the present invention. . After 24 hours, the medium in the 96-well plate was removed, 10 µl of the empty solution was added thereto, and then cultured for 1 hour at 37 ° C. in a 5% CO ₂ incubator to confirm the formation of polmament. 100 μl each of the DMSO solution was added thereto, and the activity was measured at 570 nm using a microplater.

As a result, as shown in Figure 1, compared with the control group that did not treat the chaff extract, empty reduction is inversely proportional to colorectal cancer cell lines treated with 0.5, 5, 50 and 500 mg / ml of the chaff methanol extract of the present invention It can be seen that the enemy is lowered.

That is, the chaff methanol extract of the present invention was found to have a strong anticancer activity in colorectal cancer cell line.

Example 2 Isolation of Colon Cancer Cell Line Inhibitory Substance from Methanol Extract of Chaff of the Present Invention

20 g of the rice husk methanol extract of the present invention was separated and extracted with ethyl acetate, and the extract was concentrated under reduced pressure to remove the solvent, followed by silica gel column chromatography. The eluting solvent conditions were fractionated by eluting to a chloroform-methanol (1: 1) mixed solvent while increasing the polarity by increasing the ratio of methanol from the mixed solvent of chloroform-methanol (50: 1). In addition, only the fractions showing the inhibitory activity of colon cancer cell line in each fraction were collected, and the active fractions were collected by chromatography on a Sephadex LH-20 column saturated with 100% methanol solvent. At this time, HPLC was performed using the collected active fractions as a 70% methanol solvent to obtain a pure purified colorless compound BL-1 (7 mg).

Then, the two-dimensional NMR (Fig. 2, such as the ¹ H NMR spectrum, ¹³ C NMR 1-D NMR (Fig. 1) and HMBC, HMQC, COSY spectrum such as the spectrum in order to identify the chemical structure of the obtained a colorless compound BL-1 ) Was measured.

From this result, it was confirmed that the compound structure of BL-1 is momilactone B (momilactone B).

Example 3 Growth Inhibition Activity of Colorectal Cancer Cell Line of the Compound of the Present Invention Momilactone B

In this Example, first, the anticancer activity of the compound of the present invention, momiractone B, was first used HT-29 and SW620 cell lines derived from human colorectal cancer cell lines, and cell death was measured by an empty reduction assay. The cell lines were cultured in an incubator at 37 ° C. and 5% CO ₂ using RPMI1640 medium, incubated for 72 hours in a concentration of 1.5 × 10 ⁴ cells / well in a 96-well plate, and then treated with chaff methanol extract of the present invention. . After 24, 48 and 72 hours, the medium in the 96-well plate was removed, 10 µl of the empty solution was added thereto, and then cultured for 1 hour at 37 ° C. in a 5% CO ₂ incubator to confirm the formation of polmaformation. 100 μl of DMSO solution was added to a 96-well plate and the activity was measured at 570 nm using a microplater.

As a result, as shown in FIG. 4, colorectal cancer cell lines treated with 0.5, 1, 5, and 10 mM of momiractone B showed significantly higher anticancer activity compared to the control group that did not treated with momiractone B. In particular, higher activity was observed in the SW620 cell line than the HT-29 cell line.

In addition, in order to measure the anticancer activity of the compound momiralactone B of the present invention, colony forming ability using HT-29 and SW620 cell line derived from human colorectal cancer cell line was measured. First, the cell lines were cultured in an incubator at 37 ° C. and 5% CO ₂ using RPMI1640 medium, and then cultured for 24 hours in a concentration of 1 × 10 ⁵ cells / well in a 6-well plate. Was processed. After 24 hours, the medium in the 6-well plate was removed, the cells were collected by trypsin treatment, and then cultured in the 12-well plate for 7 days. At this time, the formed colonies were fixed with 4% paraformaldehyde and stained with trypan blue solution to check their activity.

As a result, as shown in FIG. 5, colorectal cancer cell lines treated with 0.5, 1, and 5 mM of miralactone B showed significantly higher anticancer activity compared to the control group without miralactone B. The activity was higher in SW620 cell line than in 29 cell line.

In addition, a CCD986sk cell line, a fibroblast cell line derived from human skin, was used to measure normal cell line toxicity of the compound momiralactone B of the present invention, and cell death was measured by an empty reduction assay. First, the cell line was cultured in an incubator at 37 ° C. and 5% CO ₂ using RPMI1640 medium, incubated for 24 hours in a concentration of 1.5 × 10 ⁴ cells / well in a 96-well plate, followed by the present invention miralactone B Was treated. After 24 hours, the medium in the 96-well plate was removed, and 10 µl of the empty solution was added thereto, followed by incubation for 1 hour in a thermostat of 37 ° C and 5% CO ₂ to confirm the formation of polmament. 100 μl each of the DMSO solution was added thereto, and the activity was measured at 570 nm using a microplater.

As a result, as shown in Figure 6, compared to the control group not treated with mother Miractone B, low cytotoxicity was confirmed in the fibroblast cell line treated with mother Miractone B 5 and 10 mM.

As described above, the present invention confirms the anticancer activity of the chaff extract which is a by-product of agricultural processing produced in Korea, and isolates the Momiractone B compound from the chaff extract, from which the HT- derived from the human colon cancer cell line. While showing anticancer activity against the 29 and SW620 cell lines, the cytotoxicity of the normal cell line has almost no cytotoxicity, it can be applied to medicines that can selectively increase anticancer activity in solid cancers.

In addition, the present invention can produce expensive pharmaceutical products from almost discarded rice hulls, which is very useful industrially as well as economic value.

In addition, a range of modifications, changes, and substitutions may be made to the above disclosure, and in some cases only some of the features of the invention may be used. Accordingly, the appended claims should be construed broadly and in accordance with the spirit and scope of the invention.

Claims (4)

Anticancer composition containing Momilatone B as an active ingredient. The method of claim 1, Momilactone B is an anticancer composition, characterized in that extracted from chaff. The method of claim 2, The extraction is anticancer composition, characterized in that extracted with an organic solvent. The method of claim 3, wherein The organic solvent is an anticancer composition, characterized in that methanol or ethanol.

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